和解利湿方对乙醇性慢性胰腺炎腺泡细胞凋亡的影响

Effect of Hejie Lishi Formula on the Apoptosis of Pancreatic Acinar Cells in Alcohol Chronic Pancreatitis

目的:探讨和解利湿方通过抑制胰腺腺泡细胞凋亡对大鼠乙醇性慢性胰腺炎(ACP)的治疗作用。方法:将受试动物30只,随机分为2组,正常组6只,造模组24只,给予等热量的Lieber-De-Carli流质饮食,每日消毒后定量喂食14周;正常对照组6只,饲普通饮食14周;第8周末始模型组大鼠尾静脉注射给予脂多糖(LPS)剂量分别为1.5,1.5,2.0,2.5,3 mg.kg-1,1周1次,共5次,对照组给予等剂量的生理盐水;10周末造模组重分为模型组、和解利湿方高、中、低剂量(21.7,10.85,5.425 g.kg-1)治疗组,每组6只,开始ig给药,每天1次,连续给药4周。测定其组织病理变化、细胞凋亡及其相关蛋白的表达。体外观察含药血清对胰腺腺泡AR42J细胞生长和凋亡的影响。结果:与模型组比较,正常组无明显炎症症状,而和解利湿方组能减少炎症细胞浸润与胰腺腺泡细胞脱落,差异有统计学意义(P<0.05);和解利湿方高剂量组减少细胞凋亡,有显著的统计学差异(P<0.05),并能抑制半胱氨酸天冬氨酸蛋白酶(caspase)-3和caspase-9活性,增加B细胞淋巴细胞/白血病2基因(Bcl-2),降低Bcl-2相关蛋白基因(Bax)表达,其中和解利湿方高剂量组中Bcl-2/Bax是模型组的1.5倍。体外试验与含30%正常大鼠血清组比较,含30%和解利湿方药物血清明显促进胰腺腺泡细胞的生长(P<0.05);与乙醛干预并加正常大鼠血清组比较,和解利湿方含药血清组细胞凋亡率减少了32.66%。结论:高剂量的和解利湿方对ACP有一定的治疗作用,其机制可能与减少腺泡细胞凋亡、调节胰腺细胞caspase-3,caspase-9的活性及Bcl-2和Bax蛋白的表达有关。

Objective： To investigate the effects of Hejie Lishi formula on rat alcohol chronic pancreatitis （ACP） through inhibiting apoptosis of pancreatic acinar cells. Method： Thirty animals were divided into two groups. Model group of 24 animals was established by feeding Lieber-De-Carli Liquid diet after 14 weeks. Normal group of 6 animals was given general diet. The model animals were injected 1.5, 1.5, 2.0, 2.5, 3 mg .kg-1 lipopolysaccharide （LPS） into caudal vein for five times at 8 weeks, once per one week. The model group was divided into control and different concentrations （21.7, 10. 85, 5. 425 g·kg-1） of Hejie Lishi formula group at 10 weeks. Then Hejie Lishi formula was given 4 weeks and once per day by ig. After treatment for 4 weeks, all rats were executed. The pathological changes of pancreas, apoptosis and its associated proteins were detected in pancreatic tissue. Moreover, apoptosis and cell growth were observed in AR42J cells with serum contained drug treatment. Result： There were inflammatory cell infiltration, bleeding and pancreatic acinar cell shedding in control group, compared with normal. Compared to control, Hejie Lishi formula decreased inflammatory cell infiltration and pancreatic acinar cell shedding （ P 〈 O. 05 ）. The high concentration of Hejie Lishi formula reduced apoptosis （P 〈 0.05） and inhibited the activation of caspase-3 and caspase-9, increased the level of Bcl-2 and decreased the expression of Bax. The ratio of Bcl-2 to Bax in the high concentration of Hejie Lishi formula is 1.5- fold higher than control group. Compared with 30% normal rats serum, 30% containing Hejie Lishi formula serum enhanced pancreatic acinar cell growth （P 〈 0. 05）. Apoptosis was reduced 32.66% by containing Hejie Lishi formula serum. Conclusion： High concentration of Hejie Lishi formula may prevent inflammatory and injury of pancreatic ceils in ACP of rat. The mechanism may be involved in reducing apoptosis of pancreatic acinar cells through down-regulating caspase-3, -9 activities and bax expression, and up-regulating Bcl-2 expression.